Commercial Relationships: Andrea E. Hartsock, None; Jeffrey M. Costin1, Natta Sakolsatayadorn1, Stephen A. McBride2, Julian D. Purpose: To investigate polymorphic variation in the dimensions of the orbicularis oculi muscle OOM through anatomic dissection of fresh-frozen human cadavers. Methods: Skin incisions were created along a line 1 cm lateral and parallel to a line connecting the supraorbital notch SON and the infraorbital foramen IOF and from the lateral canthus to the superior border of the tragus.
The OOM was isolated using a combination of sharp and blunt dissection until each of its distal borders were identified. A metric ruler measured the superior, inferior, and lateral dimensions of the OOM from the orbital rim. Data collection included age at time of death, gender, and race.
Results: A total of 12 hemifaces from 6 fresh frozen human cadavers were dissected. All specimens were male. Cadavers were of Caucasian 5 and African 1 decent. Average age at time of death was Mean lateral OOM dimension was 2. Mean superior dimension was 1. Mean inferior dimension was 1. The average distance from the lateral orbital rim to the superior border of the tragus was 8. Increased knowledge of these dimensions has significant clinical and surgical implications, including more nuanced neurotoxin administration and surgical applications.
Commercial Relationships: Bryan R. Serial No. McBride, None; Julian D. Sorden1, Dale G. Dunn1, Peter J. Sonnentag1, Alexander J. Purpose: Rabbits are commonly used in nonclinical safety evaluation of ophthalmic drugs. To reduce the number of animals, blood samples for toxicokinetic TK and toxicity tests are often taken from the same animals. Potential catheterization-related ocular lesions occurred in previous rabbit studies. This study evaluated ocular effects of various blood collection techniques in rabbits.
On Days 1 and 7, blood was collected at 7 timepoints in an 8-hour interval via repeated hypodermic needle puncture of a medial auricular artery Group 1 , a jugular vein Group 2 , or a combination of medial auricular artery and a jugular vein Group 3 ; or via a medial auricular artery catheter Group 4. Group 5 had a catheter without a needle taped to the surface of one pinna control for Group 4, no blood collected.
Clinical signs, dermal irritation at collection sites, body weight, and anatomic pathology were assessed. Jugular venipuncture sites had the most severe skin irritation and resulted in the fewest useable samples. Findings correlated with catheter site, i. Groups 1 and 3 only had microscopic lesions at intraartery sites. Group 2 had no technique-related microscopic lesions. Conclusions: Repeated blood collection via ear artery catheter can result in foci of necrosis and inflammation in paraocular and mandibular salivary glands.
Paraocular gland lesions could confound microscopic assessment of ophthalmic drugs. A combination of jugular vein and ear artery collections is a reasonable alternative with minimal samples missed. However, a satellite group for TK sample collections is recommended for ocular toxicity studies in rabbits.
E ; Steven D. Sorden, Covance Laboratories, Inc. E ; Dale G. Dunn, Covance Laboratories Inc. E ; Peter J. Sonnentag, Covance Laboratories Inc. E ; Alexander J. Purpose: The use of supportive and spacer grafts is often necessary in eyelid reconstruction. While the use of autologous grafts minimizes tissue rejection and restores structural integrity, the size of harvested grafts and secondary wound morbidity can be limiting factors.
Furthermore, there is little information documenting the use of these grafts in combination. Methods: A retrospective chart review was performed and 6 patients were identified with cicatricial lower eyelid retraction requiring allogeneic, xenographic, or bioengineered grafts. We examined the preoperative anatomical considerations, types of procedures performed, postoperative outcomes, and complications. Results: Multiple commercially available grafts were utilized depending on the level of injury within the eyelid to restore anatomic and functional properties in all 6 patients.
The most complex reconstruction was for a patient who had an iatrogenic cicatricial entropion and eyelid retraction with hardware adhesion to an extruding orbital implant. The patient suffered from epiphora, inferior scleral show, and lagophthalmos with exposure keratopathy. None of the patients developed an infection or graft rejection. While the goal in tissue selection usually focuses on restoring form, the dynamic properties of the lower eyelid should also dictate the choice of graft. Depending on the layer s of the eyelid involved, this is currently achievable with an array of grafts used alone or in combination.
The complex anatomy and function of the lower eyelid can be safely simulated without donor site morbidity or significant complications.
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Kraus, Gregg T. Purpose: Hyperplastic persistent pupillary membranes PPM obstructing the visual axis are rare among children. Among those born with the anomaly, most regress by age one. However, persistent clinically significant obstruction of the visual axis places a child at risk of amblyopia. Therefore, medical and surgical interventions are implemented to ensure clearing of the visual axis and optimal visual development. There are many different approaches to surgical removal of PPM. We describe our specific surgical technique and the long-term visual and cosmetic results following PPM removal.
Methods: We retrospectively analyzed 10 cases of PPM removal in 6 patients. Each PPM was felt to be clinically significant based on poor visual acuity, poor retinoscope reflex, or inability to visualize the fundus. Cases of bilateral PPM underwent sequential surgery with the more visually significant membrane removed first. Surgical technique employed a clear corneal incision. A viscoelastic agent was injected beneath the pupillary strands bowing them forward. Residual synechiae were peeled from the anterior lens capsule using a Sinskey hook.
To complete the optical iridectomy, additional viscoelastic was injected beneath iris strands, which were cut using intraocular scissors. The pupillary membrane was removed with McPherson forceps. Miosis was pharmacologically induced to assess symmetric pupillary constriction. Post-operative vision and cosmesis were recorded on follow-up. Results: The PPMs were bilateral in 4 patients and unilateral in 2 patients. Age at time of surgery ranged from 2. Mean postoperative follow-up was 4.
One patient was treated for anisometropic amblyopia. No operative complications occurred. Conclusions: Several techniques have emerged for management of clinically significant PPM including nonsurgical e. This retrospective review demonstrates excellent outcomes with our technique. Potential complications of PPM removal include iris atrophy, iridodialysis, and damage to lens. We did not encounter any of these complications in our cohort. The described technique for the surgical removal of PPM represents a safe and effective treatment. We observed excellent post-operative visual acuities and cosmetic outcomes.
Commercial Relationships: Courtney L. Kraus, None; Gregg T. McNutt, Bryan R. McBride, Julian D. Purpose: To characterize the macroscopic and microscopic structure of the frontalis muscle in fresh-frozen cadavers, and specifically the frontalis dehiscence.
Uncategorized » Pathology & Laboratory Medicine | Boston University
Methods: Three fresh-frozen cadavers were marked in preparation for removal of tissue from the forehead for microscopic evaluation. Three blocks of tissue were removed at inferior, middle, and superior locations of the forehead. Dimensions of tissue blocks were 5mm x 3cm in height and width, respectively, with depth from skin to frontal periosteum. The first block of tissue was removed 1cm superior to a horizontal line drawn through the right and left supraorbital notches.
The second block was subsequently removed starting 1cm above the superior aspect of the first block and the third block removed in similar fashion 1cm from the superior aspect of the second block Fig. All blocks were marked with suture to define laterality and were submitted to pathology in formalin. Dissection was subsequently performed to expose the frontalis muscle.
Inferior incisions were made at the upper eyelids with lateral incisions made lateral to the temporal fossa so as not to disturb the frontalis muscle during reflection and removal of skin. Results: The level of frontalis dehiscence was measured at 2 cm and 3 cm in two of the three cadavers. The third cadaver was Cushingoid due to high-dose steroids and lacked macroscopically identifiable frontalis muscle. Microscopic evaluation showed a lack of muscle fibers in the midline of tissue blocks.
Conclusions: Dehiscence of the frontalis muscle observed macroscopically during gross dissection of fresh-frozen cadaveric specimens correlates with areas devoid of muscle fiber by microscopic evaluation of these same tissue samples. Clinical implications of these findings include administration of neurotoxin in the forehead, as administration in the midline forehead may not be efficacious. McNutt, None; Bryan R. Sponsel5, 1, Matthew A. Purpose: In proportion to surface area, battlefield ocular trauma is 20 to 50 times more likely than other exposed areas Sobaci et al.
Although the ability of secondary-blast effects fragments, shrapnel to cause ocular trauma is well known, it is not yet known whether ocular trauma may be induced by primary blast effects i. We hypothesized that primary, sub-lethal blast alone is sufficient to induce ocular trauma with the corollary that the level of trauma is correlated with the peak overpressure.
We tested this hypothesis using a shock-tube capable of replicating physically accurate shock-waves within a controlled environment. Methods: Porcine eyes were enucleated and set in a rigid mimic of the orbit filled with gelatin Sponsel et al. The porcine eyes were imaged via UBM and B-scan ultrasound along 12 vectors both before and after blast. Each eye was re-pressurized via intracameral injection using a 30 gauge needle with balanced salt solution and placed in a shock-tube Fig.
The eyes were then exposed to blasts of 2 ms duration with peak pressures ranging from psi. After post-blast imaging, eyes were fixed in formalin for later dissection or histopathology. Results: We observed corneal abrasion and fissuring, angle recession, circumferential iris plication into the zonule, uveal pigment forced between the axonal fasciculi of the optic nerve Fig.
Mid peripheral chorioretinal detachments, radial peripapillary retinal detachments Fig 2b , and intrinsic internal scleral delamination Fig 2c were also observed. In an extreme case, an incident pressure of 22 psi caused axial-posterior displacement of 8 mm within 67 ms and eventual evulsion of the eye from its orbit mimic on rebound. Conclusions: Primary blast in the absence of particle insult was sufficient to cause a range of significant ocular trauma. The magnitude of the incident peak-pressure was correlated with the probability of trauma occurrence and severity of damage.
The ability of primary blast overpressure to produce severe eye damage underscores the necessity of developing protective eyewear specifically designed to shield the eye and orbital structures from this source of trauma. Ahn2, 1, Milap Mehta2, Julian D. Purpose: To determine the position of a ray comprised of the anterior and posterior ethmoidal foramina as it bisects the optic canal.
Methods: A prospective case series evaluating human skulls was performed between May and June at the Cleveland Museum of Natural History. Data collected included skull demographics if available and color photographs of orbits. A ray was added to the orbital photo using Adobe Photoshop, beginning at the anterior ethmoidal foramen and extending through the posterior ethmoidal foramen and optic canal. Skulls were excluded if there was significant damage to the orbits or wear precluding the ability to easily identify the anterior and posterior ethmoidal foramina and the optic canal.
Results: A total of 5 skulls 10 orbits were available for review. There were 5 male and 0 female skulls with an average age of 48 years range, years. The APER was considered in a middle position in three orbits, superior in five orbits, and inferior in two orbits. Conclusions: The anterior and posterior ethmoidal foramina serve as critical landmarks often used when navigating towards the posterior orbit through a medial approach.
Although the APER can be a useful guide, its relationship with the optic canal varies considerably. Therefore surgeons should proceed with gentle dissection and caution when advancing towards the optic nerve using the APER alone. Commercial Relationships: Eric S. Reilly2, Brian Lund4, Rick E. Sponsel5, Randolph D. Purpose: Ocular trauma due to blast has increased dramatically in the Iraq and Afghanistan conflicts. Eye trauma has been observed in approximately one-quarter of battlefield injuries.
However, trauma from blast pressure alone, or primary blast, is not well documented as pressure is commonly accompanied by debris and acceleration of the body. Some researchers argue that primary blast cannot create significant ocular trauma, instead serious injury results from the secondary debris or body acceleration.
Recent experiments have suggested the potential for injury from high levels of blast pressure. To further evaluate this hypothesis and investigate the experimental results, high-fidelity computational models of the eye were developed and exercised. Modeling confirmed the reality of primary blast injury and allowed for identification of the physical mechanisms responsible. Methods: Numerical simulations were run in conjunction with shock tube experiments.
Sub-lethal blast levels psi; 2 ms were used in the experiments and modeling. Previous experiments focused on globe rupture, but our interest was characterizing lower but potentially serious internal injuries. High-fidelity geometrical and tissue constitutive representations were developed and implemented in both models. Results: The computational models successfully predicted the types of injury observed in the physical experiments.
Observed injuries included retinal detachment, angle recession, lens displacement, and injury to the iris and zonules. Higher levels of blast pressure were associated with higher levels of injury. However, some injury types could not be duplicated with the models. Specifically smearing of pigment into the optic nerve was observed in the experiments, but could not be duplicated with the current models. Conclusions: The numerical models confirmed the potential for ocular injury due to primary blast.
Computational results were well correlated with the physical experiments and provided invaluable insight into the mechanisms responsible for injury. The models can therefore be used in future efforts to evaluate protection schemes against primary blast. Glickman, None Support: U. Purpose: To evaluate the benefit of endonasal topical application of mitomycin-C in patients undergoing transcanalicular diode laser dacryocystorhinostomy DL-DCR. The procedure was performed under local anesthesia. In 30 patients, endonasal mitomycin-C 0.
We compared the lacrimal permeability by syringing lacrimal pathway, after 12 months of performing surgery. The results were compared using the Chi-squared test with the Yates correction. Results: The average follow-up was 12 months range 6 to 18 months. Of the patients who underwent DCR without mitomycin-C, 34 failed after a year of surgery. Only one failed of the 30 patients treated with mitomycin-C. Conclusions: In our study, the administration of topical endonasal mitomycin-C during transcanalicular diode laser dacryocystorhinostomy is associated to permeability of nasolacrimal duct.
It is a minimally invasive quick procedure yielding a high success rate. We did not find secondary effects due to application of mitomcyn-C. Commercial Relationships: Jorge E. Melanocyte morphology and pigmentation were assessed by phase-contrast microscopy. Culture purity was verified by immunostaining with melanocyte and retinal pigment epithelium RPE markers. Proliferation was measured using MTS assay. Results: Reduced oxygen conditions did not change the morphology and pigmentation of melanocytes.
Conclusions: This study demonstrated that pure long-term cultures of choroidal melanocytes could be established using physiological oxygen conditions that more closely replicates the native tissue environment. Purpose: The chorioscleral interface may be irregular in different areas of the same eye. Single point choroidal thickness measurements may be misleading.
The purpose of this study is to report the pattern and magnitude of diurnal variation of macular choroidal volume MCV measured by spectral domain optical coherence tomography SD-OCT in normal subjects. Methods: SD-OCT with enhanced depth imaging EDI and image tracking at least 49 average images using a standarized protocol was performed in 14 healthy volunteers at 2 hr intervals starting at 8 am and finishing at 6 pm. Twenty five choroidal scans centered at the fovea were performed using a raster protocol.
The choroidal thickness was segmented manually for each choroidal scan. The retinal boundary reference lines placed by the built in automated segmentation software were moved to the choroidal boundaries. This grid divided the macula into 3 circles of 1 mm central , 3 mm inner and 6 mm outer diameter. The automated software calculated the volume in each of these circles. Results: The mean age of the study subjects was The mean axial length AL was The mean diurnal variation of MCV were 0.
Conclusions: The MCV fluctuates significantly during the day in healthy subjects. Factors that influence MCV fluctuation remain to be identified. Purpose: Age-related macular degeneration AMD is a major cause of blindness. The mechanism that causes abnormal choroidal angiogenesis is not completely understood, partially due to the lack of an assay of choroidal vascular growth. Here we characterized and optimized an easily isolated, robust, quantitative and reproducible organotypic microvascular model from the choroid.
After days of incubation, the area covered by tube-like sprouts was quantified by ImageJ software and the cell types of the sprouts were analyzed by fluorescence-activated cell sorting FACS. The normalization and quantification methods for calculating the sprouting area have been standardized, and the impact of retinal pigment epithelial RPE cells, age of the animals, the type of media used for incubation and the responses of the assay to pharmacological stimulation has been characterized.
Results: The variation of vascular sprouting area is comparable between choroid samples obtained within or between animals. The sprouting area is highly reproducible between batches when normalized to controls. A semi-automated quantification macro has been developed for efficient quantification. The removal of RPE from the choroid and aging of choroid reduce the sprouting rate. Vascular endothelium growth factor VEGF promotes choroidal sprouting where 4-HDHA 4-hydroxydocosahexaenoic acid , an anti-angiogenic metabolite of omega-3 polyunsaturated fatty acid, reduces choroid sprouting dose dependently.
This method provides a new experimental tool not only for AMD studies, but also for physiological and pharmacological research related to microvascular diseases in general. Purpose: Choroidal macrophages are resident ocular immune cells capable of influencing the inflammatory environment of the outer retina and play a role in AMD pathogenesis. However, choroidal macrophages have not been characterized in detail in terms of their anatomy, distribution, behavior, and possible endogenous functions. The structure of the choroidal vasculature was simultanously visualized by the intravascular perfusion of DiI.
Perivascular macrophages associated with large choroidal arteries and primary arterioles have elongated, spindle-shaped morphologies aligned with the long axes of vessels, with secondary dendritic processes projecting into the perivascular space. Macrophages associated with smaller terminal arterioles demonstrate a more symmetric, ramified morphology and are mainly positioned within interstitial spaces between vessels and at vessel branch points.
Macrophages associated with the choriocapillaris are located only on the scleral, but not the vitreal, choriocapillaris surface and show a flattened morphology with symmetrically ramified processes. Live imaging experiments reveal that choroidal macrophages demonstrate dynamic surveying in their processes but exhibit limited cellular migration.
Conclusions: Choroidal macrophages demonstrate morphological diversity and varied associations with the choroidal vasculature at each level. Their dynamic process behavior and close vascular associations suggest that they may play functional roles in vasoregulation and vascular surveillance in the choroid.
Lutty1, Imran A. Bhutto1, Johanna M. Seddon2, D. Purpose: Mast cells MCs are effector cells of the innate immune system and are inhabitants of most mammalian choroids. They are activated by complement C5a , IgE, and many microorganisms. When activated, they degranulate, releasing a plethora of proteases, growth factors, endoglycosidases, histamine, and lipid metabolites. The purpose of this study was to determine MC numbers and their degranulation in age-related macular degeneration AMD compared to aged choroidal tissues.
Methods: Human postmortem eyes were obtained within 24 hours post mortem. Retina was excised from the eye cup and then the choroid dissected intact. The choroid was incubated for alkaline phosphatase enzyme activity yielding a blue formazan reaction product in viable blood vessels. After washing, the tissue was also incubated for nonspecific esterase activity, which labels MCs and granulocytes as published previously Lutty et al, A J Path The choroid was then partially bleached and counts of mast cells were made in at least three fields in each area of choroid.
Results: Human choroid had MCs randomly dispersed. The number of nondegranulated MCs in submacular choroid was greater in all groups than temporal equatorial choroid. Although the number of nondegranulated MCs was similar in submacular choroid between the GA subjects and controls mean mean Also, the GA subject had significantly less degranulated MCs in temporal equatorial choroid than submacular One control eye was note worthy: this had the highest mast cell counts of all eyes However, none of the mast cells were degranulated in this subject.
Conclusions: The significantly higher number of degranulated mast cells in GA submacular choroid suggests that mast cell degranulation may contribute to the pathologic decline of this tissue in GA. The proteases released in degranulation could degrade the choroidal stroma and Bruchs membrane and cause death of RPE and endothelial cells, while the cytokines released can attract macrophages.
Commercial Relationships: Gerard A. Lutty, None; Imran A. Bhutto, None; Johanna M. Purpose: The doublecortin-dsRed transgenic reporter rat was designed to analyze neurogenesis in the aged brain. Doublecortin DCX is specifically and transient active in neuronal precursors and young neurons. The aim of this study was to characterize possible DCX-dsRed positive cells in the adult rat retina and to analyze whether the DCX-dsRed rat might represent an appropriate model to study neuronal de- and regeneration in the rat eye in different pathological situations e.
Colocalization with dsRed positive cells was analyzed via confocal laser-scanning microscopy. About half of all dsRed positive cells in the INL were lacking the above mentioned markers and could not be classified yet. In the GCL the majority of dsRed positive cells showed colocalization with the ganglion cell marker Brn3a, while a minority displayed immunoreactivity for calbindin only probably representing displaced amacrine cells. A subpopulation was enmeshed by GFAP positive filaments and showed perivascular localization. Conclusions: In adult rat retina, DCX-dsRed cells in the GCL were identified as retinal ganglion cells, amacrine cells and perivascular cells, while in the INL part of dsRed positive cells represented horizontal cells.
Although the DCX-dsRed staining in the retina do not seem to mark retinal neuronal progenitor cells specifically, as seen in the brain, this model represents an useful tool to study retinal ganglion cells in pathological conditions, such as glaucoma. Purpose: Evidence of oxidative stress has been demonstrated in tissues with glaucoma damage. IOP was transiently elevated for 1 hour with an adjustable lasso around the right topically anesthetized eye of Sprague-Dawley rats.
IOP was measured before, immediately after, at the end of 1 hour treatment, and 1 hour after rest using TonoLab tonometry ICare, Finland. Results: Mean baseline IOP of Lipid peroxidation appears present within the eye following early IOP elevation. Commercial Relationships: Karen M. It is expressed in various regions of the brain and was lately also detected in retinal neurons of rat and mouse and in humans in intrinsic choroidal neurons.
Autonomic innervation is essential for many aspects of ocular homeostasis, and alarin might be involved in this autonomic control. Here we ask if alarin is present in the various autonomic ganglia supplying the eye and explore its impact in ocular innervation. Methods: Cranial autonomic ganglia of the rat i. For documentation, confocal laser scanning microscopy was used. Presence of alarin was quantified in ten non-consecutive serial sections of each ganglion and quantitative real-time PCR was applied to detect alarin mRNA expression in corresponding ganglia.
Results: Weak alarin-like immunoreactivity was detected in neurons of all cranial autonomic ganglia. Conclusions: Alarin has been described in various regions of the CNS and eye. Since it is only present in a minority of neurons of rat cranial autonomic ganglia, and since we were not able to detect alarin mRNA, we consider it of low impact on ocular autonomic innervation, at least under physiological conditions. Further investigations in other species are needed to clarify the role of alarin function in the eye.
Edward1, 2, Sami A. Purpose: We hypothesized that the extracellular matrix EM of encapsulated bleb played an important role in the hydraulic resistance of encapsulated blebs. To test this hypothesis we used immunohistochemistry to label the EM and proliferating fibroblasts in excised encapsulated blebs following Ahmed Glaucoma valve AGV implantation and compared them with control Tenons tissue.
The clinical features of the patients were reviewed retrospectively. AGV, model S2 was inserted in The mean interval between AGV insertion and valve revision was EM labeling was patchy with variable intensity in the excised tissue. Conclusions: The EM markers used in this study did not show differences in immunolabeling between excised capsules and control tissue.
It is possible that a EM markers other than those studied played an important role in increased hydraulic resistance or b EM density as reflected by the patchy distribution of EM marker label rather than averaging immunohistochemical label maybe important in predicting the hydraulic resistance of the capsule. Commercial Relationships: fatima fikri, None; Deepak P. Edward, None; Sami A. KU Leuven, Leuven, Belgium. Purpose: Matrix metalloproteinases MMPs cleave structural elements of the extracellular matrix and many molecules involved in signal transduction.
Although an involvement of MMPs in the proper development and regeneration of the optic circuit has been sporadically reported, their role in pathfinding of retinal ganglion cell RGCs axons and in tectal re innervation are still largely unknown. Therefore, elucidating the contribution of Mmps in formation of retinotectal projections in the developing and regenerating visual system of zebrafish might unravel novel molecules, able to support mammalian CNS re innervation.
Mmp2 and Mmpa double knockdown, performed with suboptimal doses of ATG MOs, showed a significantly decreased tectal area, as compared to embryos after single Mmpa knockdown. Importantly, active Mmp-2 levels were reduced in Mmp14a knockdown embryos, indicating that, also in zebrafish, Mmp14a contributes to Mmp2 activation. After ONC in adult zebrafish, preliminary data revealed Mmp downregulation during the axon outgrowth phase. However, when tectal reinnervation occurs, Mmp levels were found to be upregulated in RGC axons. Currently, we are further investigating whether both Mmps are involved in tectal reinnervation after ONC.
Conclusions: Overall, these findings suggest a functional link or coinvolvement between Mmp-2 and Mmp in RGC axonal re innervation of the OT in the developing and regenerating zebrafish brain. Purpose: Anterior segment dysgenesis ASD is a spectrum of disorders affecting the development of anterior structures of the eye, leading to vision loss including glaucoma.
As COL4A1 is present in all ocular basement membranes, and multiple tissues are affected in ASD, dissecting the primary site of pathogenesis can be difficult. Slitlamp examination was performed to assess the extent of ASD. Results: All three tissue-specific mutants developed ASD; however, the disease severity differed. Conclusions: Our results suggest that ASD in Col4a1 mutant mice largely results from a primary insult from ocular vasculature.
The mechanism of how abnormal ocular vasculature mutants lead to abnormal anterior segment development remains to be determined. Akula1, 2, Robert J. Munro1, Anne Moskowitz1, 2, Ronald M. Hansen1, 2, Toco Y. Chui3, Sanjay P. Prabhu4, 5, Anne B. Fulton1, 2. Purpose: Measure the effects of ROP on intraocular structures. Using software modified from our study of the rat eye Chui et al. Conclusions: Ocular abnormalities are increased more in ROP than in preterm birth alone. Commercial Relationships: James D. Akula, None; Robert J. Hansen, None; Toco Y. Chui, None; Sanjay P. Prabhu, None; Anne B.
Anderson, Yang Sun. Purpose: Pancreas transcription factor 1 alpha Ptf1a is involved in the differentiation of pancreas islet cells and may be involved in the differentation of cell types in other organ systems. In particular it has been shown to be expressed in the horizontal and amacrine cells during retinal zebrafish development.
Our purpose is to determine the presence of anterior segment tissues of zebrafish during embryogenesis. Methods: Transgenic Ptf1a:green fluorescent protein GFP strain zebrafish were acquired and allowed to develop for 5 days. Fish was embedded in Agarose gel and in-vivo imaging was performed. The zebrafish were scanned with 0. Results: Ptf1a is transiently expressed early in the anterior segment development of zebrafish, as well as in the horizontal and amacrine cells of retina.
Protein expression was evaluated and distribution of protein expression was confirmed by immunoblot analysis and immunohistochemistry. Conclusions: In this study we show Ptf1a expression in the anterior segment during early development in the zebrafish. Future steps involve determining differentiation pathways and resulting cell types with particular attention to the inner non-pigmented ciliary epithelium and possible implications for aqueous production. Purpose: Nanophthalmos is the leading cause of blindness in hereditary eye diseases, characterized by abnormal remodelling of scleral stroma.
The purpose of this study was to investigate the mechanism of nanophthalmos by assessing the pathologic features and the role of matrix metalloproteins MMPs in sclera. Methods: 80 eyes of 40 patients 15 patients from 8 pedigrees and 25 sporadic patients with nanophthalmos were studied between and Sclera samples were collected from 14 patients with nanophthalmos during sclerectomy, and sclera samples from eye bank eyes were collected as normal control.
Pathologic characters of the excised sclera were analyzed by immunohistochemical and electron microscopic examinations. Results: Nanophthalmos is characterized with short ocular axial length The collagen fiber bundles are irregularly arranged and separated into small fibrils, and glycogen granules were found accumulated between the twisting or fraying collagen fibrils in sclera of nanophthalmos. Conclusions: The abnormal remodelling of scleral stroma played important roles in the occurrence and development of nanophthalmos complication. Purpose: Ocular manifestations of Marfan syndrome include myopia and ectopia lentis.
Marfan syndrome is caused by mutations in fibrillin1, a glycoprotein enriched in force-bearing structures such as ciliary zonule. The use of mouse Marfan models for ocular studies has been limited because little is known about the nature of the murine zonule. The present study was designed to help fill this knowledge gap.
Antibodies against fibrillin isoforms and microfibril-associated glycoprotein-1 Magp1 were used for immunofluorescence applications, in conjunction with volume-rendering techniques. Results: Fbn2 was the dominant fibrillin expressed in the embryonic eye.
Fbn2 was transiently expressed in the vascular tunic, where it was found associated with Magp1 in microfibrils. By E Zonular fibers were evident by P1, after which Fbn2 expression declined and Fbn1 expression increased. By P30, a well-organized ciliary zonule was present. Zonular fibers projected from the posterior portion of the pars plicata to anterior, posterior, and equatorial termination points on the lens capsule.
The posterior fibers attached to a dense meshwork of radially-oriented microfibrils on the capsular surface. The microfibrils formed a micrometer-wide band encircling the lens. A spatial relationship between the zonular attachment and the transition zone of the lens epithelium was noted. In view of the known ability of microfibrils to modulate BMP signaling, the zonule could thus serve both a structural role and a role in lens growth. Finally, our data suggest a model for how the complex architecture of the zonule may arise.
Connections between the ciliary epithelium and the lens capsule form during embryogenesis, when the two tissues are in intimate contact. The characteristic fanning of zonular fibers may be the consequence of differential growth rates in lens and NPCE surface areas. Pretorius1, Yusuf Agamawi1, Julia M. Hatler1, Stephanie L. Bill3, Shuo Lin4, Lisa A. Purpose: Proper closure of the optic fissure during early embryogenesis is critical for normal eye formation.
Failure of optic fissure closure results in coloboma and related ocular defects. Mutations in several developmentally important genes are known to cause colobomas; however, the genetic etiology for most patients remains unknown. As a means to characterize novel genes and pathways that contribute to coloboma formation, we developed a method to evaluate optic fissure closure in zebrafish.
Methods: The genetic and developmental similarities to the mammalian eye make zebrafish Danio rerio an ideal model to study early vertebrate eye development. To evaluate optic fissure closure, we assayed for changes in Pax2a expression, a gene transiently expressed in the ventral optic cup prior to optic fissure closure. The quantity of green fluorescent protein GFP in the enhancer trap mpa:GFP transgenic zebrafish line was used as a proxy for Pax2a expression. As proof of principle, we tested whether sema3e, a gene expressed in ventral mesenchyme that exists between the edges of the optic fissure, affected GFP levels in the enhancer trap mpa:GFP transgenic line.
As a second test of optic fissure closure, we evaluated basement membrane dissolution by observing changes in laminin expression at the edges of the closing optic fissure. Results: Transgenic expression of GFP in the mpa:GFP enhancer trap line recapitulates endogenous Pax2a expression in the eye field, midbrain-hindbrain boundary, otic placode and pronephric mesoderm.
Quantitative three-dimensional analysis using ImageJ software revealed increased expression of Pax2a in sema3e knockdown embryos compared to uninjected controls at 48 hours post fertilization hpf , indicating aberrant optic fissure closure. Moreover, a reduction in overall eye size was observed at 48 hpf. We also observed that in sema3e knockdown zebrafish, laminin expression was increased compared to uninjected wild-type and the optic fissure edges exhibited delayed closure.
Conclusions: We demonstrate that quantitative three-dimensional analysis of the enhancer trap mpa:GFP transgenic zebrafish line can facilitate the identification and screening of candidate genes that impact optic fissure closure. Commercial Relationships: Pamela R. Hatler, None; Stephanie L. Universiity of Cincinnati, Cincinnati, OH. Purpose: Mammalian eye development involves a transient closure and re-opening of the eyelid. In humans, the upper and lower eyelids fuse to each other at week and they separate at months post fertilization. In mice, the eyelid development undergoes a similar process.
The mouse eyelid closes at embryonic day E EE Failure of eyelid closure in mice leads to an eye-open at birth EOB phenotype. We used genetic mutant mice with the EOB phenotype as models to evaluate whether failure of embryonic eyelid closure is associated with congenital eye diseases. Methods: Three mouse strains that displayed the EOB phenotypes were used for this study.
International society for developmental psychobiology 51st annual meeting abstracts 2018
The eyes of wild type and knockout mice were collected at the prenatal pre-eyelid closure E These eyes were subjected to histology and immunohistochemistry analyses. Results: Besides the eyelid closure defect, all the knockout mice had aberrant extraocular muscles EOM. Conclusions: Studies in the mouse models suggest that failure of eyelid closure may be responsible for congenital defects of extraocular muscle, and MAP3K1 and c-Jun may have additional roles in lens development.
Purpose: At embryonic day E Fetuses were collected at E Results: In the wild type E Besides the ocular surface epithelium, cJun was strongly expressed within the dermis of the eyelid, extra ocular muscles, brain and retina. Conclusions: Our results suggest that c-Jun expression in the ocular surface epithelium is essential for embryonic eyelid closure. Lacking genetic complementation between MAP3K1 and c-Jun suggests that these factors may control embryonic eyelid closure through independent mechanisms.
Onojafe1, Brian P. Purpose: Currently, the genetic networks underlying the closure of the optic fissure during vertebrate eye development are poorly understood. Failure of optic fissure closure leads to a potentially blinding congenital ocular malformation called uveal coloboma. Profiling of global gene expression during optic fissure closure has suggested a role for the C2H2 zinc finger proteins Nlz1 and Nlz2 in early eye development.
Knockdown of either gene in zebrafish using a morpholino strategy results in optic fissure closure defects, perhaps via dysregulation of the critical transcription factor, Pax2. The aim of this project was to determine the role of the Nlz2 gene in optic fissure closure in the mammalian eye. Methods: Long range PCR and southern blotting confirmed the homologous recombination of the knockout mouse construct.
Betagalactosidase staining was used to study Nlz2 gene expression in knockout mice. Histopathological and immunohistochemical stains of Nlz2 mouse embryo sections at E Fundus and slit lamp photography were employed in adult mice. Results: Nlz2 demonstrates homozygous lethality in knockout mice by E Beta-galactosidase staining of Nlz2 mice demonstrated widespread expression, particularly in the eyes, hindbrain, facial prominences, and limbs.
Gross and histopathological sections of the developing eye revealed a failure of the optic fissure to close in homozygous knockout mice by E Fundus and anterior segment evaluation of adult mice revealed no overt phenotypic differences between heterozygotes and wild-type mice. Conclusions: Knockout of Nlz2 in mice leads to a failure of the optic fissure to close, a phenotype which closely resembles that seen in human uveal coloboma. Additionally, the gene product appears to function in the fusion of the closely apposed edges.
As no direct NLZ2 sequence mutations were found in a cohort of patients with coloboma, further genetic studies will be conducted to elucidate the mechanism by which Nlz2 interacts with other genes or players causative of uveal coloboma. Onojafe, None; Brian P. Commercial Relationships: Paul G. Commercial Relationships: Lloyd P. Commercial Relationships: Elia J. Baird, Maria Schache. Purpose: The exact nature of the genetic mechanisms underlying ocular growth in myopia has not been fully established. A number of factors have been implicated through animal studies, in particular, the early growth response protein Egr-1 in mice Zenk, the avian homolog of Egr-1 in chicks.
In Egr-1 knockout mice, several other differentially expressed genes; pcdhb9, narf, ogdh and selenbp1 have also been identified. We wished to assess the association of these genes in common myopia, refraction, axial length, anterior chamber depth and corneal curvature in an Australian cohort. Myopia was defined as A total of individuals from the GEM study were used for analysis including myopic and non myopic individuals.
Tag single nucleotide polymorphisms SNPs were chosen to encompass 2kb upstream of the start codon through to 2kb downstream of the stop codon of the chosen genes. Statistical analysis used logistic and linear regression methods including age and sex as covariates. Bonferroni corrections were applied to account for multiple testing. Conclusions: A variant in the OGDH gene was shown to be associated with corneal curvature in this cohort.
This is the first report of OGDH, a gene involved in the tricarboxylic acid cycle, being associated with corneal curvature suggesting there may be a potential mitochondrial involvement in this phenotype. Commercial Relationships: Paul N. Tkatchenko3, Andrei V. Young1, 2.
The microarray raw data were processed by subtracting background noise and performing log2 transformation. Sample outliers were removed. The normalized microarray data were analyzed using ANOVA to identify differences in miRNA expression level between myopic and contralateral control eyes. Multiple members of the miRNA family b and c , c, g, h, and j , and a, b, e, and o , and three members of the cluster , and showed increased or decreased differential expressions.
MiR and the member cluster are involved in cell pluripotency maintenance. MiR, 16, and 21 are involved in growth and development regulation. Conclusions: Differential expression of miRNAs in several ocular tissues upon induction of experimental myopia in mice suggests a developmental and regulatory role in eye growth, and can potentially serve as therapeutic targets for treating myopia.
Tkatchenko, None; Andrei V. Crewther, Loretta Giummarra, Melanie J. Purpose: Myopia is the commonest visual disorder and a risk factor for most disorders leading to visual compromise, yet the aetiology remains elusive. Thus we aimed to examine the gene expression changes underlying the morphological, ultrastructural and physiological changes previously described, that suggest severe physiological stress in the form deprived chick model. Methods: Methods: Ten male hatchling chicks of which 6 were occluded to induce form-deprivation FD myopia by attaching a translucent polystyrene occluder to the periocular feathers of their right eye from day 3 - Data was exported as.
Unlike previous studies in this area we have extended our analyses to include ontological functional pathways that may be involved in these observed changes. The GSEA algorithm identified downregulation of genes associated with cell structure and integrity and genomic strain.
An increase in cell metabolism was noted, specifically, fatty acid oxidation as the main source of energy in these tissues. Conclusions: The pathways associated with the myopic pathophysiology include diminished glucose metabolism, cellular stress responses and cell volume control. All pathways are indicative of physiological stress and diminished ability to maintain retinal homeostasis after a week of form deprivation. The pathways include most genes described in previous studies. Human SNP research is also inconsistent but mostly identifies extracellular matrix genes that offer no indication of the mechanism underlying myopia progression.
Commercial Relationships: Sheila G. Frost, John T. Siegwart, Thomas T. Purpose: To examine gene expression in the choroid that may communicate retinally-generated GO and STOP signals to the sclera to control axial elongation and refractive error development. The untreated contralateral eyes served as controls. A total of 77 genes were examined by quantitative real-time PCR qPCR , including 14 genes that showed bidirectional expression in a wholetranscriptome analysis RNA-Seq of 3 animals from each group.
As shown in the figure, 35 of 77 genes showed significant regulation treated eye - control eye.
The remainder are primarily involved in extracellular matrix remodeling and some may have a role in vascular regulation or angiogenesis. From these results in choroid, it appears that many genes are involved in conveying GO and STOP signals to the sclera. It is highly unlikely that any single gene is primarily responsible for the control of axial elongation and refractive error. Frost, None; John T. Siegwart, None; Thomas T. Summers Rada3. Refractive errors and corneal biometric parameters were measured by, respectively, a modified Hartinger refractometer and a custom-made corneal videokeratography system.
Corneal 1. MannWhitney test was used to compare the changes between treated and control groups. When data from all birds were pooled for correlation analyses, the correlations between refractive-error components and mRNA expressions were generally higher in sclera than those in cornea.
Conclusions: Regional variations in mRNA expressions of the three target genes were found in corneal and scleral tissues. These results suggest that the eye shape remodeling during myopia development may be modulated by local molecular mechanism. Nathanson3, William K. Atropine, a broad-spectrum muscarinic receptor mAChR antagonist, is known to prevent myopia progression, but unpleasant side-effects limit its wider use and its mechanism is unknown. Although there is evidence for coupling of muscarinic receptor activation with production of nitric oxide NO by NO-synthase NOS in the retina Cimini et al.
On day 6, refractive error, axial length, equatorial length, and eye weight were measured. Control eyes were not affected by treatments to the goggled eyes so the interocular difference T-C; one-way ANOVA, Tukey post-hoc was taken as the measure of treatment effect. Pathways such as these i.
Nathanson, None; William K. Purpose: Dopamine is involved in the development of myopia and the exact mechanism has not been fully clarified. One of our previous study showed that dopamine was involved in the progression of spontaneous myopia in albino guinea pigs under natural visual environments, probably due to a reciprocal action of the two dopamine receptors: activation of the D1 receptor and inhibition of the D2 receptor appear to prevent myopia.
This study investigated whether this reciprocal action also plays a role in the development of form deprivation myopia FDM in pigmented guinea pigs Cavia porcellus. The refraction, corneal radius of curvature and axial components of the eye were measured in all animals prior to, at 1week and at 2 weeks of the experiments.
In contrast, the D2 receptor agonist quinpirole promoted the development of FDM with dose dependent. The dose of ng had largest promotion of FDM The dose of 10ng of quinpirole had no effect on FDM.
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Conclusions: Our findings suggest that dopaminergic system regulates FDM development through the reciprocal activities of two type dopamine receptors in pigmented guinea pigs: D1 receptor activation inhibits, while D2 receptor activation promotes myopia. Samuels1, Timothy W. Corson1, 2. Purpose: Retinoblastoma is the most common intraocular tumor in children, often causing blindness. Orthotopic xenograft models are a critical tool for studying new therapeutic methods.
A successful xenograft model has been generated by intravitreal injection of newborn rats with a bioluminescent derivative of the Y79 human retinoblastoma cell line. Although this model is powerful, screening panels of xenografts from multiple cell lines would be valuable for assessing inter-individual responses to novel therapeutics. We evaluated whether optical coherence tomography OCT could be used to identify successful xenografts of other, non-engineered retinoblastoma cell lines and characterize xenograft growth patterns.
The Micron III rodent imaging system was used to obtain fundus photographs and OCT images regularly over the course of 4 weeks to document xenograft development. The xenografts produced both small and large tumors that were typically dense, highly vascularized, and had well- defined edges.
Successful nonfluorescent xenografts had similar morphologic characteristics and growth patterns to those from the Y79 line on OCT imaging.
- University of Veterinary Medicine Vienna - Research portal.
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Conclusions: OCT imaging of retinoblastoma orthotopic xenografts is a novel way to spatially analyze and follow tumor growth in vivo. When tumors were not always readily evident on brightfield imaging, OCT proved to be a valuable tool to help identify hard-to-see, nonfluorescent tumors generated with these non-engineered retinoblastoma cell lines. This approach will enable rapid screening of additional cell lines in the future as well as quantitative spatial analysis of response to therapeutics. Ali4, Geeta K. Prasad Eye Institute, Hyderabad, India. Purpose: Gene expression studies in Cancer provide an insight into the global functioning of tumor and their pathways.
Since, CD44 and CD90 expression was absent in Y79 cell line, we used CD to sort the cells and analysed for various stem cell assays. This study highlights gene expression signature specific to putative cancer stem cells in Y79 cell line. Total RNA was isolated and quantified. The data was analyzed and normalized using GeneSpring and Lowess algorithm. Data validation was done using semi quantitative PCR. Pathway and interaction studies were analyzed using GeneMania and String database.
Pathways upregulated in CD cells were involved in cell migration 3. Further studies to validate these pathways are warranted to understand the role of cancer stem cells in Retinoblastoma. Ali, None; Geeta K. Purpose: Retinoblastoma RB , a primary pediatric intraocular tumor. Several novel molecular strategies are being developed for the clinical management of RB. Results: In the 21 tumor tissues analysed, miRb was detected in In-vitro silencing of these miRNAs in RB cells resulted in the reduction of cell proliferation and in colony formation, through the induction of apoptosis confirmed by Annexin V assay.
Conclusions: The aberrant expression of miRbfamily in primary retinoblastoma implicates its role in RB tumorigenesis. Volkert2, Maria I. Hennig3,4, Thomas J. Nieland3,4, Daniel M. Fass3,4, Patricia F. Kendra Iskander , a surgical resident conducting research in the Remick Lab on a T32 Fellowship, was awarded a resident scholarship to attend the Society of Thoracic Surgeons meeting in Ft. The meeting should be good for networking and professional development. Nikolajczyk Lab: Dr. Panchenko Lab: Dr. Henerdon: "Cytotoxicity of amyloidogenic immunoglobulin light chains on human podocytes in culture".
This project is a collaborative effort with Dr. Lawreen Connors and Dr. Daniel G. Remick was selected by the graduate students of Wayne State to be the visiting professor. Nancy S. Title: Exosomal miRNA profiling in prefrontal cortices of patients diagnosed with bipolar disorder and schizophrenia. Vaickus, Jacqueline C.
Bouchard, Dominic Beal, William W. The SFARI program seeks to improve the diagnosis and treatment of autism spectrum disorders by funding, catalyzing, and driving innovative research of the greatest quality and relevance. This grant grew out of work by one of the former pathology graduate students, Bryan Belikoff. The grant was funded for 4 years and the work will be done in collaboration with Dr. Sitkovsky at Northeastern University Dr. Tomolonis1, Daniel J. Pollak, MD3 and Joel M. Henderson, MD, PhD 1. Tomolonis1 , Laura M. Connors , PhD2 and Joel M. The provisional patent also has been filed for this novel technology.
Kroll also presented at a roundtable on Laboratory Management On behalf of Dr. Christensen , Claudio Morera, and Julia Y. This work involved collecting samples from coastal residents to query relationships between blood levels of environmental pollutants and the astonishing rate of type 2 diabetes in the poor of Ecuador.
The project included presentation of Dr. Daniel Remic k chaired the advocacy presentations at the Association of Pathology Chairs annual meeting. The invited speaker was Dora Hughes, M. Stephen Hammond and the MGH digital pathology group. Attendees were treated to informative, interesting tours of the facility and refreshments to celebrate the seamless transition. Congratulations for a job well done! The goal of the project is to assess relationships between industrialization, industrial pollution, and the astonishing rates of obesity and type 2 diabetes among the local Ameridians native population.
The project is in year 2 of a ten year pan, and anyone interested in joining the group next year is encouraged to get in touch with Dr. Nikolajczyk directly; bnikol bu. This Committee reports to the Secretary of HHS on major issues regarding the safety and availability of blood in the country, and also on organ transplantation. Heather Cohen, 5th year PhD candidate was informed that her abstract has been accepted for poster presentation during the 12th International Workshop on Scleroderma Research at Trinity College Cambridge, United Kingdom July , Every year newly board-certified Pathologists must earn at least 10 of these credits.
Eosinophilic myocarditis in hypereosinophilic syndrome. American Journal of Hematology ; Computed tomography of granulomatous pneumonia with oxalosis in an American alligator Alligator mississippiensis associated with Metarhizium anisopliae var. Journal of Zoo and Wildlife Medicine in press.
Robbins Award for The Stanley L. Robbins Award for Excellence in Teaching is given each year at graduation to the outstanding educator at the School of Medicine. The award was established in recognition of the excellence in teaching and devotion to students exemplified by Dr. Stanley L. This award serves to acknowledge the importance of teaching skills and commitment to students and education at BUSM. This is a once in a lifetime award! Michael H. Roehrl, M. Barbara Slacks says: Congratulations to the award winners, and thanks to everyone who submitted abstracts for the poster session.
This year May 31st will be the written part of the exam, June 8th and 9th the oral portion will take place.